Showing papers in "Mycorrhiza in 2005"
TL;DR: The main conclusions are that the species structure of the ECM community and metabolic activity of each morphotype change depending on the season, temperature and soil moisture, and a number of morphotypes are more abundant and active in winter than in summer.
Abstract: This work was aimed at understanding how the functional diversity of ectomycorrhizas (ECM) is driven by environmental factors and how it adapts to the structure of the forest stand. Superficial fine roots were sampled 21 times during an entire year in two adjacent plots (no thinning and strong thinning) of a mature beech (Fagus silvatica) forest. Individual ectomycorrhizal root tips were morphologically characterised and the symbiotic fungi were molecularly identified. ECM were also tested for dehydrogenase and acid phosphatase activities, and soil moisture and temperature were recorded. The results provide a description of ECM community dynamics over a whole year in the two stands. The main conclusions are threefold: (1) the species structure of the ECM community and metabolic activity of each morphotype change depending on the season, temperature and soil moisture, and a number of morphotypes are more abundant and active in winter than in summer, (2) the silviculture treatment (strong thinning) modifies the ectomycorrhizal community structure, and (3) the overall function of the ECM community results from the individual time pattern and specialisation of each morphotype.
219 citations
TL;DR: In this paper, the authors investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques.
Abstract: We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H2O2 accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl3, intracellular accumulation of H2O2 was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H2O2 was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.
185 citations
TL;DR: GmosPT showed an expression pattern similar to GiPT, the Glomus intraradices PT gene, since its transcript was more abundant in the extraradical mycelium treated with micromolar Pi levels, which opens new questions about the role and functioning of high-affinity PT in AM fungi.
Abstract: Arbuscular mycorrhizal (AM) fungi have long been shown to successfully contribute to phosphate uptake by plant roots. The first step of the fungus-mediated uptake is carried out by fungal membrane Pi transporters (PT) that transfer Pi from the soil into the extraradical hyphae. In the present work we report the identification and characterisation of a PT gene from Glomus mosseae, an AM fungus important for natural and agricultural ecosystems. Degenerate primers and rapid amplification of cDNA ends-polymerase chain reaction (PCR) allowed us to obtain a sequence (GmosPT) showing a highly significant similarity with GiPT and GvPT, the only two other PT genes already isolated from AM fungi. Reverse transcriptase-PCR experiments were carried out to study GmosPT expression profiles in structures corresponding to different fungal life stages (quiescent and germinated sporocarps, intraradical and extraradical hyphae) and in extra- and intraradical hyphae exposed to high and low Pi concentrations. GmosPT showed an expression pattern similar to GiPT, the Glomus intraradices PT gene, since its transcript was more abundant in the extraradical mycelium treated with micromolar Pi levels. In addition, the intraradical mycelium also showed a significant GmosPT expression level that was independent from external Pi concentrations. This finding opens new questions about the role and functioning of high-affinity PT in AM fungi.
156 citations
TL;DR: There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems, and in pine, the highest level of Colonisation was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls.
Abstract: Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.
149 citations
TL;DR: Some effects of colonization by an arbuscular mycorrhizal (AM) fungus on the biomass and arsenate uptake of an As hyperaccumulator, Pteris vittata, are reported for the first time.
Abstract: We report for the first time some effects of colonization by an arbuscular mycorrhizal (AM) fungus (Glomus mosseae) on the biomass and arsenate uptake of an As hyperaccumulator, Pteris vittata. Two arsenic levels (0 and 300 mg As kg−1) were applied to an already contaminated soil in pots with two compartments for plant and hyphal growth in a glasshouse experiment. Arsenic application had little or no effect on mycorrhizal colonization, which was about 50% of root length. Mycorrhizal colonization increased frond dry matter yield, lowered the root/frond weight ratio, and decreased frond As concentration by 33–38%. Nevertheless, transfer of As to fronds showed a 43% increase with mycorrhizal colonization at the higher soil As level. Frond As concentrations reached about 1.6 g kg−1 (dry matter basis) in non-mycorrhizal plants in the As-amended soil. Mycorrhizal colonization elevated root P concentration at both soil As levels and mycorrhizal plants had higher P/As ratios in both fronds and roots than did non-mycorrhizal controls.
125 citations
TL;DR: Seasonal variation in population attributes of arbuscular mycorrhizal fungi over 2 years in four sites of temperate grasslands of the Argentinean Flooding Pampas is studied, suggesting differences among AM fungus species with respect to the seasonality of sporulation.
Abstract: We studied seasonal variation in population attributes of arbuscular mycorrhizal (AM) fungi over 2 years in four sites of temperate grasslands of the Argentinean Flooding Pampas. The sites represent a wide range of soil conditions, hydrologic gradients, and floristic composition. Lotus glaber, a perennial herbaceous legume naturalised in the Flooding Pampas, was dominant at the four plant community sites. Its roots were highly colonised by AM fungi. Temporal variations in spore density, spore type, AM root colonisation, floristic composition and soil chemical characteristics occurred in each site and were different among sites. The duration of flooding had no effect on spore density but depressed AM root colonisation. Eleven different types of spores were recognized and four were identified. Two species dominated at the four sites: Glomus fasciculatum and Glomus intraradices. Spore density was highest in summer (dry season) and lowest in winter (wet season) with intermediate values in autumn and spring. Colonisation of L. glaber roots was highest in summer or spring and lowest in winter or autumn. The relative density of G. fasciculatum and G. intraradices versus Glomus sp. and Acaulospora sp. had distinctive seasonal peaks. These seasonal peaks occurred at all four sites, suggesting differences among AM fungus species with respect to the seasonality of sporulation. Spore density and AM root colonisation when measured at any one time were poorly related to each other. However, spore density was significantly correlated with root colonisation 3 months before, suggesting that high colonisation in one season precedes high sporulation in the next season.
122 citations
TL;DR: Evidence is provided that arbuscular mycorrhiza can be much more effective than kinetin applications in protecting mungbean plants against the detrimental effects of salt water.
Abstract: Increasing use of saline water in irrigation can markedly change the physical and chemical properties of soil. An experiment was carried out to investigate the interaction between the mycorrhizal fungus Glomus clarum, isolated from a saline soil, and kinetin on the growth and physiology of mungbean plants irrigated with different dilutions of seawater (0, 10, 20, and 30%). The growth, chlorophyll concentration and sugar content of mycorrhizal plants was greater than that of non-mycorrhizal plants under all conditions (with or without seawater). The dry weight of both mycorrhizal and non-mycorrhizal mungbean plants irrigated with 10% seawater was significantly increased by treatment with kinetin. The mycorrhizal symbiosis increased root:shoot dry weight ratio, concentrations of N, P, K, Ca and Mg, plant height, protein content, nitrogen or phosphorus-use efficiencies, and root nitrogenase, acid or alkaline phosphatase activities of seawater-irrigated mungbean plants, with little or no effect of kinetin. Kinetin treatment generally decreased chlorophyll concentration and sugar content in mycorrhizal plants as well as Na/N, Na/P Na/K, Na/Ca and Na/Mg ratios. Root colonization by G. clarum was increased by irrigation with seawater, and kinetin had no consistent effect on fungal development in roots. This study provides evidence that arbuscular mycorrhiza can be much more effective than kinetin applications in protecting mungbean plants against the detrimental effects of salt water.
121 citations
TL;DR: This study for the first time documented that oribatid mites feed on certain ectomycorrhizal fungi, with O. tibialis being most selective.
Abstract: The coexistence of a large number of soil animals without extensive niche differentiation is one of the great riddles in soil biology. The main aim of this study was to explore the importance of partitioning of food resources for the high diversity of micro-arthropods in soil. In addition, we investigated if ectomycorrhizal fungi are preferentially consumed compared to saprotrophic fungi. Until today, ectomycorrhizal fungi have never been tested as potential food resource for oribatid mites. We offered six ectomycorrhizal fungi [Amanita muscaria (L.) Hook., Boletus badius (Fr.) Fr., Cenococcum geophilum Fr., Laccaria laccata (Scop.) Fr., Paxillus involutus (Batsch) Fr. and Piloderma croceum J. Erikss. & Hjortstam], one ericoid mycorrhizal fungus [Hymenoscyphus ericae (D.J. Read) Korf & Kernan] and three saprotrophic fungi [Agrocybe gibberosa (Fr.) Fayod, Alternaria alternata (Fr.) Keissl. and Mortierella ramanniana (A. Moller) Linnem.] simultaneously to each of the mainly mycophagous oribatid mite species Carabodes femoralis (Nicolet), Nothrus silvestris Nicolet and Oribatula tibialis Nicolet. The ericoid mycorrhizal fungus H. ericae and the ectomycorrhizal fungus B. badius were preferentially consumed by each oribatid mite species. However, feeding preferences differed significantly between the three species, with O. tibialis being most selective. This study for the first time documented that oribatid mites feed on certain ectomycorrhizal fungi.
120 citations
TL;DR: Roots of plants in the long established meadow and heath communities adjacent to the forefront and the high alpine community all had one or another of the colonization types, with DS and AM predominating.
Abstract: Lyman glacier in the North Cascades Mountains of Washington has a subalpine forefront characterized by a well-developed terminal moraine, inconspicuous successional moraines, fluting, and outwash. These deposits were depleted of symbiotic fungi when first exposed but colonized by them over time after exposure. Four major groups of plant species in this system are (1) mycorrhiza-independent or facultative mycotrophic, (2) dependent on arbuscular mycorrhizae (AM) (3) dependent on ericoid mycorrhiza (ERM) or ectomycorrhizae (EM), and (4) colonized by dark-septate (DS) endophytes. We hypothesized that availability of mycorrhizal propagules was related to the success of mycorrhiza-dependent plants in colonizing new substrates in naturally evolved ecosystems. To test this hypothesis roots samples of 66 plant species were examined for mycorrhizal colonization. The plants were sampled from communities at increasing distances from the glacier terminus to compare the newest communities with successively older ones. Long established, secondary successional dry meadow communities adjacent to the glacier forefront, and nearby high alpine communities were sampled for comparison. DS were common on most plant species on the forefront. Nonmycorrhizal plants predominated in the earlier successional sites, whereas the proportion of mycorrhizal plants generally increased with age of community. AM were present, mostly at low levels, and nearly absent in two sites of the forefront. ERM were present in all species of Ericaceae sampled, and EM in all species of Pinaceae and Salicaceae. Roots of plants in the long established meadow and heath communities adjacent to the forefront and the high alpine community all had one or another of the colonization types, with DS and AM predominating.
119 citations
TL;DR: Arbuscular mycorrhizal fungal spore communities were surveyed in a long-term field fertilization experiment in Switzerland, where different amounts of phosphorus were applied to soil, and no evidence has been acquired for effect of P fertilization on spore density, composition, and diversity of AMF in both the field soil and in trap cultures.
Abstract: Arbuscular mycorrhizal fungal (AMF) spore communities were surveyed in a long-term field fertilization experiment in Switzerland, where different amounts of phosphorus (P) were applied to soil. Plots receiving no P as well as plots systematically fertilized in excess to plant needs for 31 years were used to test the hypothesis that application of P fertilizer changes the composition and diversity of AMF communities. AMF spores were isolated from the field soil, identified, and counted so as to quantify the effect of P fertilization on AMF spore density, composition, and diversity. Trap cultures were established from field soil with four host plants (sunflower, leek, maize, and Crotalaria grahamiana), and the spore communities were then analyzed in substrate samples from the pots. Altogether, nine AMF species were detected in the soil. No evidence has been acquired for effect of P fertilization on spore density, composition, and diversity of AMF in both the field soil and in trap cultures. On the other hand, we observed strong effect of crop plant species on spore densities in the soil, the values being lowest under rapeseed and highest under Phacelia tanacetifolia covercrop. The identity of plant species in trap pots also significantly affected composition and diversity of associated AMF communities, probably due to preferential establishment of symbiosis between certain plant and AMF species. AMF spore communities under mycorrhizal host plants (wheat and Phacelia in the fields and four host plant species in trap pots) were dominated by a single AMF species, Glomus intraradices. This resulted in exceptionally low AMF spore diversity that seems to be linked to high clay content of the soil.
106 citations
TL;DR: Investigation of the mechanism and the extent to which AM can prevent drought damages and whether physiological analyses reveal enhanced drought tolerance of an economically important plant such as the rose found proline content in leaves proved to be an unsuitable marker for AM-induced drought tolerance.
Abstract: Micropropagated rose plants (Rosa hybrida L., cv. New Dawn) were inoculated with the arbuscular mycorrhizal (AM) fungus Glomus intraradices (Schenk and Smith) and subjected to different drought regimens. The dual objectives of these experiments were to investigate the mechanism and the extent to which AM can prevent drought damages and whether physiological analyses reveal enhanced drought tolerance of an economically important plant such as the rose. In a long-term drought experiment with four different water regimens, visual scoring of wilt symptoms affirmed that AM in a selected host-symbiont combination increased plant performance. This effect was mostly expressed if moderate drought stress was constantly applied over a long period. In a short-term experiment in which severe drought stress was implemented and plants were allowed to recover after 4 or 9 days, no visual differences between mycorrhizal and non-mycorrhizal roses were observed. Therefore, the early physiological steps conferring drought tolerance were prone to investigation. Proline content in leaves proved to be an unsuitable marker for AM-induced drought tolerance, whereas analysis of chlorophyll a fluorescence using the JIP test (collecting stress-induced changes of the polyphasic O-J-I-P fluorescence kinetics in a non-destructive tissue screening) was more explanatory. Parameters derived from this test could describe the extent of foliar stress response and help to differentiate physiological mechanisms of stress tolerance. AM led to a more intense electron flow and a higher productive photosynthetic activity at several sites of the photosynthetic electron transport chain. A K step, known as a stress indicator of general character, appeared in the fluorescence transient only in drought-stressed non-mycorrhizal plants; conversely, the data elucidate a stabilising effect of AM on the oxygen-evolving complex at the donor site of photosystem (PS) II and at the electron-transport chain between PS II and PS I. If drought stress intensity was reduced by a prolonged and milder drying phase, these significant tolerance features were less pronounced or missing, indicating a possible threshold level for mycorrhizal tolerance induction.
TL;DR: Finds contribute knowledge about the above- and below-ground ascomycete community in a ponderosa pine forest after a low intensity fire and four of the six RFLP types clustered with two genera of the Pezizales, Wilcoxina and Geopora.
Abstract: Post-fire Pezizales fruit commonly in many forest types after fire. The objectives of this study were to determine which Pezizales appeared as sporocarps after a prescribed fire in the Blue Mountains of eastern Oregon, and whether species of Pezizales formed mycorrhizas on ponderosa pine, whether or not they were detected from sporocarps. Forty-two sporocarp collections in five genera (Anthracobia, Morchella, Peziza, Scutellinia, Tricharina) of post-fire Pezizales produced ten restriction fragment length polymorphism (RFLP) types. We found no root tips colonized by species of post-fire Pezizales fruiting at our site. However, 15% (6/39) of the RFLP types obtained from mycorrhizal roots within 32 soil cores were ascomycetes. Phylogenetic analyses of the 18S nuclear ribosomal DNA gene indicated that four of the six RFLP types clustered with two genera of the Pezizales, Wilcoxina and Geopora. Subsequent analyses indicated that two of these mycobionts were probably Wilcoxina rehmii, one Geopora cooperi, and one Geopora sp. The identities of two types were not successfully determined with PCR-based methods. Results contribute knowledge about the above- and below-ground ascomycete community in a ponderosa pine forest after a low intensity fire.
TL;DR: Results indicate that although heavy metal chelation pathways contribute to Cd stress responses in pea, they may not make a major contribution toCd tolerance strategies operating in the arbuscular mycorrhizal symbiosis.
Abstract: Molecular responses to cadmium (Cd) stress were studied in mycorrhizal and non-mycorrhizal Pisum sativum L. cv. Frisson inoculated with Glomus intraradices. Biomass decreases caused by the heavy metal were significantly less in mycorrhizal than in non-mycorrhizal plants. Real-time reverse transcriptase-polymerase chain reaction showed that genes implicated in pathways of Cd detoxification varied in response to mycorrhiza development or Cd application. Expression of a metallothionein-encoding gene increased strongly in roots of Cd-treated non-mycorrhizal plants. Genes encoding gamma-glutamylcysteine synthetase and glutathione (GSH) synthetase, responsible for the synthesis of the phytochelatin (PC) precursor GSH, were activated by Cd in mycorrhizal and non-mycorrhizal plants. Cd stress decreased accumulation of GSH/homoglutathione (hGSH) and increased thiol groups in pea roots, whether mycorrhizal or not, suggesting synthesis of PCs and/or homophytochelatins. An hGSH synthetase gene, involved in hGSH synthesis, did not respond to Cd alone but was activated by mycorrhizal development in the presence of Cd. Transcript levels of a glutathione reductase gene were only increased in non-mycorrhizal roots treated with Cd. Studies of three stress-related genes showed that a heat-shock protein gene was activated in mycorrhizal roots or by Cd and chitinase gene transcripts increased under Cd stress to a greater extent in mycorrhizal roots, whilst a chalcone isomerase gene was only up-regulated by Cd. Results indicate that although heavy metal chelation pathways contribute to Cd stress responses in pea, they may not make a major contribution to Cd tolerance strategies operating in the arbuscular mycorrhizal symbiosis.
TL;DR: The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi.
Abstract: Effects of humic substances (humic acid or fulvic soil extract) or saprophytic microorganisms (Paecilomyces lilacinus and an unidentified actinomycete) on growth of mycelium and mycorrhiza formation by Glomus claroideum BEG23 were studied in a hydroponic system. Humic substances stimulated root colonization and production of extraradical mycelium by the mycorrhizal fungus. Both humic and fulvic acids tended to decrease populations of culturable bacteria and fungi in the cultivation system, indicating a moderately antibiotic activity. The addition of saprophytic microorganisms able to use humic substances to the cultivation system further stimulated the development of the mycorrhizal fungus. However, stimulation of G. claroideum was also observed when the saprophytic microorganisms were heat-killed, suggesting that their effect was not linked to a specific action on humic substances. The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi.
TL;DR: Increased density predominantly influenced plant responses by affecting whether or not carbon (photosynthate) or phosphorus limited plant growth, by exacerbating carbon limitation, high density reduced the benefit/cost ratio of mycorrhizas and minimized their effects.
Abstract: We examined the effects of arbuscular mycorrhizas (AM), phosphorus fertilization, intraspecific density, and their interaction, on the growth, phosphorus uptake, and root morphology of three facultative mycotrophic crops (Capsicum annuum, Zea mays, and Cucurbita pepo). Plants were grown in pots with or without AM at three densities and four phosphorus availabilities for 10 weeks. AM colonization, plant weight, and shoot phosphorus concentration were measured at harvest. Root morphology was assessed for C. annuum and Z. mays. Phosphorus fertilization reduced but did not eliminate AM colonization of all species. AM, phosphorus, and density interacted significantly to modify growth of C. annuum and C. pepo such that increased density and phosphorus diminished beneficial effects of AM. Increased density reduced positive effects of AM on C. annuum and C. pepo shoot phosphorus concentrations. AM altered both Z. mays and C. annuum root morphology in ways that complemented potential phosphorus uptake by mycorrhizas, but increased density and phosphorus diminished these effects. We infer that increased density predominantly influenced plant responses by affecting whether or not carbon (photosynthate) or phosphorus limited plant growth. By exacerbating carbon limitation, high density reduced the benefit/cost ratio of mycorrhizas and minimized their effects.
TL;DR: The results of this study suggest the use of adapted AMF as inoculants for phytorestoration of alkaline anthropogenic-stressed sediments in order to promote plant establishment and growth under highly alkaline conditions.
Abstract: A field survey of the arbuscular mycorrhizal status of herbaceous plant species was conducted in a highly alkaline anthropogenic sediment resulting from the disposal of waste from an acetylene and polyvinyl chloride factory. Most plant species found at the site were mycorrhizal and the dominant mycotrophic plant species was Conyza bilbaoana. Fungal species richness was assessed by identification of spores extracted from the sediment and from continuously propagated trap pot cultures. All of the six species of arbuscular mycorrhizal fungi (AMF) found were from the genus Glomus. Glomus intraradices and G. mosseae were found in field-collected sediment samples and also occurred most frequently in trap cultures. To test the symbiotic effectiveness of these two fungi, seedlings of C. bilbaoana were inoculated with either native G. intraradices BEG163 or G. mosseae BEG198 and non-native G. intraradices BEG75 or G. mosseae BEG25 isolates in sterile and non-sterile sediment collected from the study site. All four isolates were able to colonise C. bilbaoana. However, AMF native to the target sediments were generally more effective than the non-native fungi in promoting plant establishment and growth under highly alkaline conditions. The non-native G. intraradices was, however, more effective than the non-native G. mosseae. The results of this study suggest the use of adapted AMF as inoculants for phytorestoration of alkaline anthropogenic-stressed sediments.
TL;DR: There was no significant variation in the relative abundance of the ECM morphotypes between seasons, but ECM community species richness was highest in autumn and lowest in summer, and the percentage of mycorrhizal tips reached a maximum in winter, with its minimum in autumn.
Abstract: Two study plots, burned and control, were established in autumn 1998 in a Quercus ilex forest located in northern Spain, part of which had been affected by a low intensity fire in 1994. Soil samples for ectomycorrhizae (ECM) were taken over a 3-year period in each study plot in spring, summer, autumn and winter. ECM morphotypes were identified and the relative abundance of each morphotype in each soil sample calculated, along with species richness, Shannon diversity index and percentage of mycorrhization in each soil sample. The relative abundance of certain ECM morphotypes differed between burned and control plots, and the percentage of mycorrhizal tips was significantly lower in the burned than in the control plot. Nevertheless, there were no significant differences in the diversity, species richness or species composition of the ECM community in the burned and control plots. The dominant ECM morphotypes in both stands were Cenococcum geophilum and several thelephoroid fungi. Sphaerosporella brunnea and Pisolithus tinctorius thrived especially in the burned plot, whereas three ectomycorrhizal morphotypes assigned to the genus Hebeloma were especially abundant in the control plot. There was no significant variation in the relative abundance of the ECM morphotypes between seasons, but ECM community species richness was highest in autumn and lowest in summer. The percentage of mycorrhizal tips reached a maximum in winter, with its minimum in autumn. Collection of samples over the 3-year period also enabled us to detect a significant increase in percentage of ECM colonisation in the burned stand over time.
TL;DR: The fact that certain tree species, above all the Cupuliferae, do not nourish themselves independently in the soil but regularly establish a symbiosis with fungal mycelium over their entire root system is concerns, which presupposes knowledge about the nature and nutrition of plants not heretofore even slightly suspected by science.
Abstract: To promote the possibility of truffle cultivation in the Kingdom of Prussia, His Excellency, the Minister of Agriculture, Domains and Forestry, commissioned me to approach thematter systematically. I was to begin with scientific studies on the conditions of occurrence and development of these fungi. Certain facts had already been established through observations and experience. For example, true truffles occur only with living trees, and in the Prussian truffle districts these investigations had established a strong relationship between truffle occurrence and particular tree species: beech, hornbeam and oak. Above all else was the union of Elaphomyces mycelium with pine roots, as recognized by Rees (Sitzungsber. D. physik.-med. Soc. zu Erlangen, 10 May 1880). From the outset these facts pointed the research towards determining whether the true truffles also establish a connection of the mycelium with living tree roots. As this communication will show, the question must begin much farther back, because it presupposes knowledge about the nature and nutrition of plants not heretofore even slightly suspected by science. This shall be nearly the only topic of my present paper. It concerns the fact that certain tree species, above all the Cupuliferae, do not nourish themselves independently in the soil but regularly establish a symbiosis with fungal mycelium over their entire root system. This mycelium performs a “wet nurse” function and performs the entire nourishment of the tree from the soil. Surprising though this may sound, it is solidly based on the scope of my research. When one examines feeder rootlets in the soil—the root system’s end branches representing the actual organs of nutrient uptake—of any of our native oaks, beech, hornbeam, hazel or chestnut, it is evident they are generally composed of two disparate components: a core, representing the actual tree root, and an organically united mantle of fungal hyphae. This fungal mantle completely encloses the rootlet, forming a continuous cover even over the growing tip. It grows along with the root tip and behaves in every respect as an organically united, peripheral tissue belonging to the root. The entire structure is neither tree root nor fungus alone but resembles the lichen thallus, a union of two different organisms into a single, morphological organ. It can be appropriately designated as a “fungus-root” or “mycorrhiza.” Translation from German of “Über die auf Wurzelsymbiose beruhende Ernährung gewisser Bäume durch unterirdische Pilze” from Berichte der Deutschen Botanischen Gesellschaft (1885) 3:128–145, revised from an earlier translation in Molina R (ed) Proceedings of the 6th North American Conference on Mycorrhizae (1985) Translator: James M. Trappe (e-mail: trappej@onid.orst.edu, Tel.: +1-541-7378593, Fax: +1-541-7371393), Department of Forest Science, Oregon State University, Corvallis, OR 97331–5752, USA Translator’s note: This is not a literal translation, because the style of nineteenth century German scientific writing often does not translate comfortably into twenty-first century English. For example, the original German title of Frank’s paper word-for-word in English would read, “About the on-root-symbiosis-depending nutrition of certain trees through underground fungi.” In adjusting syntax in the translation, I have paid particular attention to preserving Frank’s original intent and meaning. Frank used the family name “Cupuliferae,” which was interpreted in the nineteenth century to include the present Betulaceae and Fagaceae, or sometimes only the Fagaceae, but has long since been discarded. Frank used it in this paper generally in reference to Carpinus, Corylus, Castanea, Fagus and Quercus spp.). In deriving the term “mycorrhiza” from the Greek μυκοριζα, Frank transliterated it with a single “r”. Transliterating the Greek ρ (rho) into Latin letters ideally requires that the “r” be doubled and followed by an “h” in certain compound words to make the meaning and derivation clear, as discussed byW. T. Stearn [Botanical Latin, 4th Addition (1992) Timber Press, Portland, Ore., p 261). However, many authors, including Linnaeus and Frank, omitted the additional “r.” For purposes of the science of mycorrhizae, it is trivial: whether with one “r” or two, our communication is equally effective. Similarly, whether one uses the Greek plural “mycorrhiza,” the Latin plural “mycorrhizae,” the English plural “mycorrhizas,” or the plural endings of other languages is of no consequence. “Rhiza” is feminine in Greek; Frank retained that gender in German, using German feminine case endings. To reflect his usage, I have used the English rather than Latinized possessive and plural endings in this translation.
TL;DR: The disturbed sites showed less plant diversification, with the community of arbuscular mycorrhizal fungi being quantitative and qualitatively affected by disturbance.
Abstract: The occurrence of arbuscular mycorrhizal fungi (AMF) in a copper mining area was investigated Soil samples were collected from six sites at the Mineracao Caraiba, Bahia State, northeastern Brazil, comprising: (1) a site that receives the waste product; (2) a site that receives low grade deposits; (3) the interface between the caatinga and site 1; (4) the surroundings of the industrial area; (5) the site for extracting topsoil for land filling; (6) the preserved caatinga Thirty-two plant species were identified around the collection locations Trap cultures were maintained in the greenhouse for 3 months, using bahia grass ( Paspalum notatum Flugge) as the host plant Spores were extracted from soil and 21 AMF species (15 Glomus and one of each of Acaulospora, Archaeospora, Entrophospora, Gigaspora , Paraglomus and Scutellospora) were identified In site 1, plants or AMF were not found during the dry season Site 6, with native vegetation, had the highest number of plants and AMF species The disturbed sites showed less plant diversification, with the community of AMF being quantitative and qualitatively affected by disturbance
TL;DR: This work demonstrates that there is a high correspondence between basidiomycete diversity detected by molecular analysis of root tips and soil samples, irrespective of the soil fraction being analysed.
Abstract: Molecular methods based on soil DNA extracts are increasingly being used to study the fungal diversity of ectomycorrhizal (EM) fungal communities in soil. Contrary to EM root tip identification, the use of molecular methods enables identification of extramatrical mycelia in soil. To compare fungal diversity as determined by root tip identification and mycelial identification, six soil samples were analysed. Root tips were extracted from the six samples and after amplification, the basidiomycete diversity on the root tips was analysed by denaturing gradient gel electrophoresis (DGGE). The soil from the six samples was sieved, total soil DNA was extracted and after amplification, the basidiomycete diversity in the soil fractions was analysed by DGGE. Fourteen different bands were excised from the DGGE gel and sequenced; fungal taxon names could be assigned to eight bands. Out of a total of 14 fungal taxa detected in soil, 11 fungal taxa were found on root tips, of which seven were EM fungal taxa. To examine whether the sieving treatment would affect EM species diversity, two different sieve mesh sizes were used and in addition, the organic soil fraction was analysed separately. DGGE analysis showed no differences in banding pattern for the different soil fractions. The organic fraction gave the highest DGGE band intensities. This work demonstrates that there is a high correspondence between basidiomycete diversity detected by molecular analysis of root tips and soil samples, irrespective of the soil fraction being analysed.
TL;DR: This work analyzed mycorrhizal data from an oak woodland from two perspectives: the phytocentric view using trees as nodes and fungi as links and the mycocentric View, suggesting that certain fungus species may act as hubs with frequent connections to the network.
Abstract: The concept of a common mycorrhizal network implies that the arrangement of plants and mycorrhizal fungi in a community shares properties with other networks. A network is a system of nodes connected by links. Here we apply network theory to mycorrhizas to determine whether the architecture of a potential common mycorrhizal network is random or scale-free. We analyzed mycorrhizal data from an oak woodland from two perspectives: the phytocentric view using trees as nodes and fungi as links and the mycocentric view using fungi as nodes and trees as links. From the phytocentric perspective, the distribution of potential mycorrhizal links, as measured by the number of ectomycorrhizal morphotypes on trees of Quercus garryana, was random with a short tail, implying that all the individuals of this species are more or less equal in linking to fungi in a potential network. From the mycocentric perspective, however, the distribution of plant links to fungi was scale-free, suggesting that certain fungus species may act as hubs with frequent connections to the network. Parallels exist between social networks and mycorrhizas that suggest future lines of study on mycorrhizal networks.
TL;DR: Fungal hyphae were able to respond to host-derived signals by reorienting their growth towards roots and to perceive chemotropic signals at a distance of at least 910 μm from roots, eventually establishing mycorrhizal symbiosis.
Abstract: In this work, we report the occurrence of chemotropism in the arbuscular mycorrhizal (AM) fungus Glomus mosseae. Fungal hyphae were able to respond to host-derived signals by reorienting their growth towards roots and to perceive chemotropic signals at a distance of at least 910 μm from roots. In order to reach the source of chemotropic signals, hyphal tips crossed interposed membranes emerging within 1 mm from roots, eventually establishing mycorrhizal symbiosis. The specificity of chemotropic growth was evidenced by hyphal growth reorientation and membrane penetration occurring only in experimental systems set up with host plants. Since pre-symbiotic growth is a critical stage in the life cycle of obligate AM fungal symbionts, chemotropic guidance may represent an important mechanism functional to host root location, appressorium formation and symbiosis establishment.
TL;DR: It is found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P, and inoculation may be the only option for management of the symbiosis.
Abstract: A field study was done to assess the potential benefit of arbuscular mycorrhizal (AM) inoculation of elite strawberry plants on plant multiplication, under typical strawberry nursery conditions and, in particular, high soil P fertility (Mehlich-3 extractible P=498 mg kg−1). Commercially in vitro propagated elite plants of five cultivars (‘Chambly,’ ‘Glooscap,’ ‘Joliette,’ ‘Kent,’ and ‘Sweet Charlie’) were transplanted in noninoculated growth substrate or in substrate inoculated with Glomus intraradices or with a mixture of species (G. intraradices, Glomus mosseae, and Glomus etunicatum) at the acclimation stage and were grown for 6 weeks before transplantation in the field. We found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P. Inoculated mother plants produced about 25% fewer daughter plants than the control in Chambly (P=0.03), and Glooscap produced about 50% more (P=0.008) daughter plants when inoculated with G. intraradices, while the productivity of other cultivars was not significantly decreased. Daughter plant shoot mass was not affected by treatments, but their roots had lower, higher, or similar mass, depending on the cultivar–inoculum combination. Root mass was unrelated to plant number. The average level of AM colonization of daughter plants produced by noninoculated mother plants did not exceed 2%, whereas plants produced from inoculated mothers had over 10% of their root length colonized 7 weeks after transplantation of mother plants and ∼6% after 14 weeks (harvest), suggesting that the AM fungi brought into the field by inoculated mother plants had established and spread up to the daughter plants. The host or nonhost nature of the crop species preceding strawberry plant production (barley or buckwheat) had no effect on soil mycorrhizal potential, on mother plant productivity, or on daughter plant mycorrhizal development. Thus, in soil excessively rich in P, inoculation may be the only option for management of the symbiosis.
TL;DR: The results indicate that the morphology of colonization is strongly influenced by the selection of fungi to colonize the host plant from among those in the soil environment.
Abstract: Colonization by arbuscular mycorrhizal (AM) fungi was investigated in cucumber (Cucumis sativus), tomato (Lycopersicon esculentum) and Clethra barbinervis (Ericales) grown in field-collected soil known from previous studies to generate Paris-type arbuscular mycorrhizae in C. barbinervis. Spores of Paraglomus, Acaulospora, Glomus, and Gigaspora were found in the soil. Formation of hyphal coils and arbusculate coils of Paris-type mycorrhizae and of arbuscules of Arum-type mycorrhizae in roots raised in this soil in the growth chamber were compared with the detection of DNA of AM fungi from the same root systems using Glomales-specific primers. Only Paris-type mycorrhizae with extensive arbusculate coils developed in C. barbinervis, but cucumber and tomato developed both Paris- and Arum-types in the same root systems. Glomaceae and Archaeosporaceae and/or Paraglomaceae were detected strongly in the DNA from both cucumber and tomato roots, in which Arum-type mycorrhizae were observed. In contrast, DNA of Glomaceae was detected more sparingly in C. barbinervis, in which Paris-type mycorrhizae dominated. Acaulosporaceae and Gigasporaceae were strongly detected in the DNA from both C. barbinervis and tomato, whereas they were more weakly detected in cucumber. These results indicate that the morphology of colonization is strongly influenced by the selection of fungi to colonize the host plant from among those in the soil environment.
TL;DR: This work demonstrates that caution must be taken when using this molecular approach to determine in planta AM fungal diversity if non-sequence based methods such as terminal restriction fragment length polymorphisms, denaturing gradient gel electrophoresis, or temperature gradient gel electrolysis are used.
Abstract: An increasingly popular approach used to identify arbuscular mycorrhizal (AM) fungi in planta is to amplify a portion of AM fungal small subunit ribosomal DNA (SSU-rDNA) from whole root DNA extractions using the primer pair AM1-NS31, followed by cloning and sequencing. We used this approach to study the AM fungal community composition of three common oak-woodland plant species: a grass (Cynosurus echinatus), blue oak (Quercus douglasii), and a forb (Torilis arvensis). Significant diversity of AM fungi were found in the roots of C. echinatus, which is consistent with previous studies demonstrating a high degree of AM fungal diversity from the roots of various hosts. In contrast, clones from Q. douglasii and T. arvensis were primarily from non-AM fungi of diverse origins within the Ascomycota and Basidiomycota. This work demonstrates that caution must be taken when using this molecular approach to determine in planta AM fungal diversity if non-sequence based methods such as terminal restriction fragment length polymorphisms, denaturing gradient gel electrophoresis, or temperature gradient gel electrophoresis are used.
TL;DR: The trends revealed in this study corroborate earlier studies suggesting a predominance of multiple host fungi in mixed communities of EM plants and support the prediction that western hemlock can associate with fungi in Douglas-fir EM networks.
Abstract: We investigated ectomycorrhizal (EM) fungi in forest stands containing both early successional Douglas-fir and late successional western hemlock at two points in the typical stand development by identifying EM fungi from roots of Douglas-fir and western hemlock in mixed stands. In an early seral stage forest, EM roots of western hemlock seedlings and intermingling 40-year-old Douglas-fir were sampled. In a late seral stage forest, EM roots of trees of both species were sampled in a 400-year-old stand. We use molecular approaches to identify the symbionts from field samples in this descriptive study. In the early seral stage study, >95% of the western hemlock root tips by biomass were colonized by fungi also colonizing Douglas-fir roots. This result supports the prediction that western hemlock can associate with fungi in Douglas-fir EM networks. In the same study, fungi specific to Douglas-fir colonized 14% of its EM root tips. In the late seral stage study, 14% of the western hemlock root tips were colonized by fungi also observed in association with Douglas-fir, a result strongly influenced by sampling issues and likely represents a conservative estimate of multiple host fungi in this old growth setting. Fungi specific to Douglas-fir colonized 25% of its root tip biomass in the old growth study, in tight coralloid clusters within five of the 24 soil samples. The trends revealed in this study corroborate earlier studies suggesting a predominance of multiple host fungi in mixed communities of EM plants. The role of host-specific fungi in these stands remains unclear.
TL;DR: The reliance of AM colonisation on stem C declined with a decrease in colonisation, providing more C for the refilling of stem carbohydrate reserves and shoot growth, and AM roots were more efficient at P utilisation.
Abstract: Arbuscular mycorrhizal (AM) C-costs in grapevines were investigated. Dormant vines rely on stored C for initial growth. Therefore AM colonisation costs would compete with plant growth for available C reserves. One-year-old grapevines, colonised with Glomus etunicatum (Becker and Gerdemann), were cultivated under glasshouse conditions. The C-economy and P utilisation of the symbiosis were sequentially analysed. AM colonisation, during the 0–67 day growth period, used more stem C relative to root C, which resulted in lower shoot growth. The decline in AM colonisation during the period of 67–119 days coincided with stem C replenishment and higher shoot growth. Construction costs of AM plants and root C allocation increased with root P uptake. The efficiency of P utilisation was lower in AM roots. The reliance of AM colonisation on stem C declined with a decrease in colonisation, providing more C for the refilling of stem carbohydrate reserves and shoot growth. Once established, the AM symbiosis increased P uptake at the expense of refilling of root C reserves. Although higher root C allocation increased plant construction costs, AM roots were more efficient at P utilisation.
TL;DR: Agrobacterium tumefaciens-mediated gene transfer should facilitate future development of insertional mutagenesis, targeted gene disruption and RNA interference technology in L. A. bicolor.
Abstract: The development of an efficient transformation system is required to alter the expression of symbiosis-regulated genes and to develop insertional mutagenesis in the ectomycorrhizal basidiomycete Laccaria bicolor S238N. Vegetative mycelium of this fungus was transformed by Agrobacterium tumefaciens-mediated gene transfer. The selection marker was the hygromycin resistance gene of Escherichia coli (hph) under the control of the gpd promoter from Agaricus bisporus and the CaMV 35S terminator as part of the T-DNA. PCR amplification of hph and Southern blot analyses showed that the genome of the hygromycin-resistant transformants contained the cassette. The latter proved mostly single copy and random integration of part of the transgene into the fungal genome. A. tumefaciens-mediated gene transfer should facilitate future development of insertional mutagenesis, targeted gene disruption and RNA interference technology in L. bicolor.
TL;DR: The revolution in thinking about plant and fungal evolution, ecology and physiology generated by Frank is still in the process of acceptance by much of the scientific community, 120 years and tens of thousands of scientific papers since he coined the term “mycorrhiza”.
Abstract: A. B. Frank's observations and hypotheses about mycorrhizae in 1885 flew in the face of conventional thinking of the time. He reported that what we now term ectomycorrhizae were widespread on root systems of many woody plant species in a great diversity of habitats and soils. He hypothesized that mycorrhizae represent a pervasive mutualistic symbiosis in which fungus and host nutritionally rely on each other; that the fungus extracts nutrients from both mineral soil and humus and translocates them to the tree host; and that the tree, in turn, nourishes the fungus. Initially opposed by much of the scientific community, nearly all of Frank's major hypotheses have since been unequivocally demonstrated, although many decades were required to achieve conclusive evidence. Nonetheless, the revolution in thinking about plant and fungal evolution, ecology and physiology generated by Frank is still in the process of acceptance by much of the scientific community, 120 years and tens of thousands of scientific papers since he coined the term "mycorrhiza". The reasons for this extraordinary lag time in themselves present an intriguing research subject.
TL;DR: The mycorrhizal crops strawberry and caraway maintained RMEDW most effectively, while the values were lower in the non-host crops buckwheat, turnip rape and fiddleneck, and the numbers of AM spores detected in soil were considerably greater during 3 years of strawberry cultivation.
Abstract: Mycotrophy of previous crops has been shown to have an impact on arbuscular mycorrhizal fungi (AMF), and the growth and productivity of succeeding crops. We studied the impact of 3 years of cultivation of eight crops with different degrees of mycotrophy, including mycorrhizal (strawberry, rye, timothy, onion, caraway) and non-mycorrhizal (turnip rape, buckwheat, fiddleneck) hosts, as well as the impact of peat amendment, on the effectiveness, amount and diversity of indigenous AMF. A field experiment having a split-plot design with peat amendment as the main plot, crop cultivation as a sub-plot and three replications, was carried out on silt clay mineral soil in 1999-2001. A well-humified dark peat was applied immediately before establishment of the field experiment. Each year, the relative mycorrhizal effectiveness of soil collected in September, in terms of shoot dry weight (RME(DW)), was determined in a bioassay. In the 3rd year of the experiment, AMF spores were also extracted and identified from the field soil. Expressed as the mean of 3 years of cropping in unamended soil, the mycorrhizal crops strawberry and caraway maintained RME(DW) most effectively, while the values were lower in the non-host crops buckwheat, turnip rape and fiddleneck. In addition, the numbers of AM spores detected in soil were considerably greater during 3 years of strawberry cultivation. In soil under caraway, there were high numbers of AM spores compared to the other crops. In soil amended with peat, the situation was in some cases opposite of that of unamended soil; RME(DW) was highest in rye and onion and lowest in strawberry and caraway. The reasons behind the negative impact of peat on mycorrhizal effectiveness in strawberry soil may be due to the microbiological properties of peat. The importance of including mycotrophic species in crop rotations for maintaining high soil quality and for increasing yields of subsequent crops is discussed.